Kanno M, Kobayashi S, Tokuhisa T, Takei I, Shinohara N, Taniguchi M
J Exp Med. 1981 Nov 1;154(5):1290-304. doi: 10.1084/jem.154.5.1290.
The B cell hybridomas producing monoclonal antibodies (E10, D7, F4, H6, and D4) were established by the fusion of P3U1 or NS-1 murine myeloma cell lines and spleen cells of B10.A(5R) mice hyperimmunized with mitomycin C-treated B10.A(3R) spleen and thymus cells. Two types of monoclonal antibodies specific for the products controlled by a gene in the I-Jb subregion of the H-2 complex were characterized: one specific for the private type of I-Jb determinant, the other recognizing the cross-reactive determinant between the I-Jb and I-Jd products. By using these monoclonal reagents, the I-J-encoded product on the antigen-specific suppressor T cells was found to be expressed on their soluble suppressor factors. Furthermore, the I-Jb products were successfully detected not only on the T cell hybridoma with suppressor activity specific for keyhole limpet hemocyanin (KLH), but also on KLH-primed suppressor T cells enriched by antigen-coated petri dishes and concanavalin A-induced thymocyte blasts of C57BL/6 mice by complement-dependent cytotoxic assays and membrane fluorescence techniques.
通过将P3U1或NS-1鼠骨髓瘤细胞系与经丝裂霉素C处理的B10.A(3R)脾细胞和胸腺细胞免疫的B10.A(5R)小鼠的脾细胞融合,建立了产生单克隆抗体(E10、D7、F4、H6和D4)的B细胞杂交瘤。鉴定了两种对由H-2复合体I-Jb亚区中的一个基因控制的产物具有特异性的单克隆抗体:一种对I-Jb决定簇的私有类型具有特异性,另一种识别I-Jb和I-Jd产物之间的交叉反应决定簇。通过使用这些单克隆试剂,发现抗原特异性抑制性T细胞上由I-J编码的产物在其可溶性抑制因子上表达。此外,不仅在对钥孔血蓝蛋白(KLH)具有特异性抑制活性的T细胞杂交瘤上成功检测到I-Jb产物,而且通过补体依赖性细胞毒性测定和膜荧光技术,在通过抗原包被的培养皿富集的KLH致敏的抑制性T细胞以及C57BL/6小鼠的伴刀豆球蛋白A诱导的胸腺细胞母细胞上也成功检测到了I-Jb产物。